Chalcone structure fluorescence dye for embryonic stem cell probe

Abstract

The invention includes a compound represented by the following structural formula: wherein is described herein. The compounds of the invention are useful in staining embryonic stem cells.

What is claimed is: 
     
       1. A compound represented by the following structural formula: 
       
         
           
           
               
               
           
         
         or a salt thereof, wherein: 
         R is (C 1 -C 16 )alkyl, (C 2 -C 16 )alkenyl, (C 2 -C 16 )alkynyl, carbocyclyl or heterocyclyl, each of which is optionally substituted with one or more groups independently selected from the following: halo, NO 2 , —NHCO(C 1 -C 6 )alkyl, —OCO(C 1 -C 6 )alkyl, —COO(C 1 -C 6 )alkyl, (C 1 -C 10 )alkyl, halo(C 1 -C 10 )alkyl, (C 1 -C 10 )alkoxy, —S(C 1 -C 6 )alkyl, (C 3 -C 9 )cycloalkyl, aryl, and phenoxy, wherein each phenoxy or aryl are further optionally substituted with one or more groups selected from halo, NO 2 , (C 1 -C 10 )alkyl, halo(C 1 -C 10 )alkyl, and (C 1 -C 10 )alkoxy. 
       
     
     
       2. The compound of  claim 1 , or a salt thereof, wherein R is (C 1 -C 16 )alkyl. 
     
     
       3. The compound of  claim 2 , or a salt thereof, wherein R is (C 1 -C 11 )alkyl, optionally substituted with one, two or three groups independently selected from chloro, bromo, methyl, ethyl, methoxy, —COOCH 3 , —OCOCH 3 , cyclopentyl, phenyl, and phenoxy, wherein the phenyl or phenoxy is optionally and independently substituted with one, two, three, four, or five groups independently selected from fluoro, chloro, and methoxy. 
     
     
       4. The compound of  claim 1 , or a salt thereof, where the compound is represented by the following structural formulas: 
       
         
           
           
               
               
           
         
       
     
     
       5. The compound of  claim 1 , or a salt thereof, wherein R is C 2 -C 16 )alkenyl. 
     
     
       6. The compound of  claim 5 , or a salt thereof, wherein R is —CH═CH 2 , —CH 2 CH 2 C═CH 2 , (CH 2 ) 8 C═CH 2 , optionally substituted with one or more substituents selected from methyl and phenyl, wherein the phenyl is optionally substituted with chloro. 
     
     
       7. The compound of  claim 6 , or a salt thereof, wherein the compound is represented by the following structural formula: 
       
         
           
           
               
               
           
         
         or a salt thereof. 
       
     
     
       8. The compound of  claim 1 , or a salt thereof, wherein R is carbocyclyl. 
     
     
       9. The compound of  claim 8 , or a salt thereof, wherein R is an optionally substituted aryl. 
     
     
       10. The compound of  claim 9 , or a salt thereof, wherein R is an optionally substituted phenyl. 
     
     
       11. The compound of  claim 10 , or a salt thereof, wherein R is phenyl, optionally substituted with one, two, three, four or five groups independently selected from halo, NO 2 , —NHCO(C 1 -C 6 )alkyl, —OCO(C 1 -C 6 )alkyl, (C 1 -C 10 )alkyl, halo(C 1 -C 4 )alkyl, (C 1 -C 10 )alkoxy, and phenyl. 
     
     
       12. The compound of  claim 11 , or a salt thereof, wherein R is phenyl, optionally substituted with one, two, three, four or five groups independently selected from fluoro, chloro, bromo, iodo, —NO 2 , —NHCOCH 3 , —OCOCH 3 , methyl, ethyl, propyl, isopropyl, butyl, pentyl, hexyl, heptyl, nonyl, decyl, —CF 3 , —CH 2 Cl, methoxy, ethoxy, butoxy, hexoxy, heptoxy, and phenyl. 
     
     
       13. The compound of  claim 9 , or a salt thereof, wherein R is naphthyl, optionally substituted with one, two, three, four or five groups independently selected from halo, NO 2 , —NHCO(C 1 -C 6 )alkyl, —OCO(C 1 -C 6 )alkyl, (C 1 -C 10 )alkyl, halo(C 1 -C 4 )alkyl, (C 1 -C 10 )alkoxy, and phenyl. 
     
     
       14. The compound of  claim 13 , or a salt thereof, wherein R is naphthyl, optionally substituted with ethoxy. 
     
     
       15. The compound of  claim 8 , or a salt thereof, wherein R is (C 3 -C 9 )cycloalkyl, optionally substituted with one or two groups independently selected from halo, NO 2 , —NHCO(C 1 -C 6 )alkyl, —OCO(C 1 -C 6 )alkyl, (C 1 -C 10 )alkyl, halo(C 1 -C 4 )alkyl, (C 1 -C 10 )alkoxy, and phenyl. 
     
     
       16. The compound of  claim 15 , or a salt thereof, wherein R is cyclohexyl, cyclopentyl, cyclobutyl, or cyclopropyl. 
     
     
       17. The compound of  claim 8 , or a salt thereof, wherein R is (C 3 -C 9 )cycloalkenyl, optionally substituted with one or two groups independently selected from halo, NO 2 , —NHCO(C 1 -C 6 )alkyl, —OCO(C 1 -C 6 )alkyl, (C 1 -C 10 )alkyl, halo(C 1 -C 4 )alkyl, (C 1 -C 10 )alkoxy, and phenyl. 
     
     
       18. The compound of  claim 17 , or a salt thereof, wherein R is cyclohexenyl. 
     
     
       19. The compound of  claim 1 , or a salt thereof, wherein R is heterocyclyl. 
     
     
       20. The compound of  claim 19 , or a salt thereof, wherein R is 
       
         
           
           
               
               
           
         
       
     
     
       21. The compound of  claim 19 , or a salt thereof, wherein R is optionally substituted heteroaryl. 
     
     
       22. The compound of  claim 21 , or a salt thereof, wherein R is optionally substituted heteroaryl selected from benzo[b]thiophene, furanyl, pyrrole, isoxazole, thiophene, pyridine, and pyrimidine. 
     
     
       23. The compound of  claim 22 , or a salt thereof, wherein R is selected from 
       
         
           
           
               
               
           
         
         wherein each R is optionally substituted with one, two, or three groups independently selected from fluoro, chloro, bromo, —NO 2 , methyl, methoxy, —SCH 3 , and phenyl, wherein the phenyl is optionally substituted with one, two or three groups independently selected from fluoro, chloro, and NO 2 . 
       
     
     
       24. A method for the preferential staining of stem cells, comprising incubating a cell population comprising stem cells with a compound of Formula (I) or a salt thereof: 
       
         
           
           
               
               
           
         
         wherein: 
         R is (C 1 -C 16 )alkyl, (C 2 -C 16 )alkenyl, (C 2 -C 16 )alkynyl, carbocyclyl or heterocyclyl, each of which is optionally substituted with one or more groups independently selected from the following: halo, NO 2 , —NHCO(C 1 -C 6 )alkyl, —OCO(C 1 -C 6 )alkyl, —COO(C 1 -C 6 )alkyl, (C 1 -C 10 )alkyl, halo(C 1 -C 10 )alkyl, (C 1 -C 10 )alkoxy, —S(C 1 -C 6 )alkyl, (C 3 -C 9 )cycloalkyl, aryl, and phenoxy, wherein each phenoxy or aryl are further optionally substituted with one or more groups selected from halo, NO 2 , (C 1 -C 10 )alkyl, halo(C 1 -C 10 )alkyl, and (C 1 -C 10 )alkoxy; 
         for a period of time sufficient to stain stem cells, wherein stem cells are stained with greater intensity by fluorescence than any other cell types. 
       
     
     
       25. The method of  claim 24 , further comprising sorting the stained stem cells from the remaining cell population. 
     
     
       26. The method of  claim 25 , wherein the sorting is Fluorescence Activated Cell Sorting (FACS). 
     
     
       27. The method of  claim 24 , wherein the compound of Formula (I) is: 
       
         
           
           
               
               
           
         
         or a salt thereof. 
       
     
     
       28. The method of  claim 24 , wherein the stem cells are embryonic stem cells. 
     
     
       29. The method of  claim 24 , wherein the stem cells are induced pluripotent stem cells. 
     
     
       30. A method of visualizing a stem cell, comprising:
 a) contacting a cell population with a compound of formula (I) or a salt thereof: 
 
       
         
           
           
               
               
           
         
         
           wherein: 
           R is (C 1 -C 16 )alkyl, (C 2 -C 16 )alkenyl, (C 2 -C 16 )alkynyl, carbocyclyl or heterocyclyl, each of which is optionally substituted with one or more groups independently selected from the following: halo, NO 2 , —NHCO(C 1 -C 6 )alkyl, —OCO(C 1 -C 6 )alkyl, —COO(C 1 -C 6 )alkyl, (C 1 -C 10 )alkyl, halo(C 1 -C 10 )alkyl, (C 1 -C 10 )alkoxy, —S(C 1 -C 6 )alkyl, (C 3 -C 9 )cycloalkyl, aryl, and phenoxy, wherein each phenoxy or aryl are further optionally substituted with one or more groups selected from halo, NO 2 , (C 1 -C 10 )alkyl, halo(C 1 -C 10 )alkyl, and (C 1 -C 10 )alkoxy; 
           to form an incubation media; 
         
         b) incubating the incubation media of step (a) for a period of time sufficient to stain the stem cells; and 
         c) visualizing the stained cells of step (b) with fluorescence microscopy to visualize a stem cell. 
       
     
     
       31. The method of  claim 30 , wherein the stem cell is an embryonic stem cell.